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Article

Continuous Cell Lines from the European Biting Midge Culicoides nubeculosus (Meigen, 1830)

1
Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, 146 Brownlow Hill, Liverpool L3 5RF, UK
2
UMR1161 Virologie, INRAE, Ecole Nationale Vétérinaire d’Alfort, ANSES, Université Paris-Est, 94700 Maisons-Alfort, France
3
The Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK
4
School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington Campus, Sutton Bonington LE12 5RD, UK
*
Author to whom correspondence should be addressed.
Microorganisms 2020, 8(6), 825; https://doi.org/10.3390/microorganisms8060825
Received: 12 May 2020 / Revised: 25 May 2020 / Accepted: 28 May 2020 / Published: 30 May 2020
(This article belongs to the Special Issue Bluetongue Virus)
Culicoides biting midges (Diptera: Ceratopogonidae) transmit arboviruses of veterinary or medical importance, including bluetongue virus (BTV) and Schmallenberg virus, as well as causing severe irritation to livestock and humans. Arthropod cell lines are essential laboratory research tools for the isolation and propagation of vector-borne pathogens and the investigation of host-vector-pathogen interactions. Here we report the establishment of two continuous cell lines, CNE/LULS44 and CNE/LULS47, from embryos of Culicoides nubeculosus, a midge distributed throughout the Western Palearctic region. Species origin of the cultured cells was confirmed by polymerase chain reaction (PCR) amplification and sequencing of a fragment of the cytochrome oxidase 1 gene, and the absence of bacterial contamination was confirmed by bacterial 16S rRNA PCR. Both lines have been successfully cryopreserved and resuscitated. The majority of cells examined in both lines had the expected diploid chromosome number of 2n = 6. Transmission electron microscopy of CNE/LULS44 cells revealed the presence of large mitochondria within cells of a diverse population, while arrays of virus-like particles were not seen. CNE/LULS44 cells supported replication of a strain of BTV serotype 1, but not of a strain of serotype 26 which is not known to be insect-transmitted. These new cell lines will expand the scope of research on Culicoides-borne pathogens. View Full-Text
Keywords: Culicoides; Ceratopogonidae; Monoculicoides; cell line; vector; midge; bluetongue virus; orbivirus; virus replication Culicoides; Ceratopogonidae; Monoculicoides; cell line; vector; midge; bluetongue virus; orbivirus; virus replication
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MDPI and ACS Style

Bell-Sakyi, L.; Mohd Jaafar, F.; Monsion, B.; Luu, L.; Denison, E.; Carpenter, S.; Attoui, H.; Mertens, P.P.C. Continuous Cell Lines from the European Biting Midge Culicoides nubeculosus (Meigen, 1830). Microorganisms 2020, 8, 825. https://doi.org/10.3390/microorganisms8060825

AMA Style

Bell-Sakyi L, Mohd Jaafar F, Monsion B, Luu L, Denison E, Carpenter S, Attoui H, Mertens PPC. Continuous Cell Lines from the European Biting Midge Culicoides nubeculosus (Meigen, 1830). Microorganisms. 2020; 8(6):825. https://doi.org/10.3390/microorganisms8060825

Chicago/Turabian Style

Bell-Sakyi, Lesley, Fauziah Mohd Jaafar, Baptiste Monsion, Lisa Luu, Eric Denison, Simon Carpenter, Houssam Attoui, and Peter P.C. Mertens 2020. "Continuous Cell Lines from the European Biting Midge Culicoides nubeculosus (Meigen, 1830)" Microorganisms 8, no. 6: 825. https://doi.org/10.3390/microorganisms8060825

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